RIPAb+? G3BP1

• 产品编号：merck-m-03-180      品牌：millipore       原厂货号：03-180
• 产品分类：抗体 > 一抗 > 蛋白特异性一抗
• 应用分类：

描述：

Description:RIPAb+™ G3BP1 - RIP Validated Antibody and Primer Set

Overview:RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation.
G3BP1 (Ras-GTPase activating protein SH3-domain binding protein 1) directly associates with SH3 domains of GTPase activating proteins (GAPs). G3BP1, along with G3BP2, has been shown to bind to the C-terminus of p53, playing a crucial role in its activity (Kim, 2007). Furthermore, the G3BPs have two traditional RNA binding motifs.

Background Information:G3BP1 (Ras-GTPase activating protein SH3-domain binding protein 1) directly associates with SH3 domains of GTPase activating proteins (GAPs). It was first discovered by its binding to RasGAP. G3BPs have been shown to be involved in a number of mitogenic signaling pathways. G3BP1 is also known to be over expressed in many human cancers (Kim, 2007) and its expression appears to be inversely correlated with PTEN expression (Kim, 2007). Interestingly, G3BP1, along with G3BP2, has been shown to bind to the C-terminus of p53, playing a crucial role in its activity (Kim, 2007). Furthermore, the G3BPs have two traditional RNA binding motifs.

Presentation

Anti-G3BP1 (mouse monoclonal),. One vial containing 50 µg of protein G purified mouse monoclonal IgG1ĸ in buffer containing 0.1 M Tris-glycine, 0.150 M NaCl, pH 7.4, and 0.05% sodium azide before the addition of 30% glycerol. Store at -20°C .
Normal Mouse IgG, . One vial containing 125 µg of mouse IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C.
RIP Primers, c-myc 3' UTR. One vial containing 75 μL of 5 μM of each primer specific for human c-myc 3' UTR. Store at -20°C.
FOR: CAC AAA CTT GAA CAG CTA CGG
REV: GGT GAT TGC TCA GGA CAT TTC 

Key Applications

• Western Blotting
• RNA Binding Protein Immunoprecipitation (RIP)

原厂资料：

Description:RIPAb+™ G3BP1 - RIP Validated Antibody and Primer Set

Overview:RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation.
G3BP1 (Ras-GTPase activating protein SH3-domain binding protein 1) directly associates with SH3 domains of GTPase activating proteins (GAPs). G3BP1, along with G3BP2, has been shown to bind to the C-terminus of p53, playing a crucial role in its activity (Kim, 2007). Furthermore, the G3BPs have two traditional RNA binding motifs.

Background Information:G3BP1 (Ras-GTPase activating protein SH3-domain binding protein 1) directly associates with SH3 domains of GTPase activating proteins (GAPs). It was first discovered by its binding to RasGAP. G3BPs have been shown to be involved in a number of mitogenic signaling pathways. G3BP1 is also known to be over expressed in many human cancers (Kim, 2007) and its expression appears to be inversely correlated with PTEN expression (Kim, 2007). Interestingly, G3BP1, along with G3BP2, has been shown to bind to the C-terminus of p53, playing a crucial role in its activity (Kim, 2007). Furthermore, the G3BPs have two traditional RNA binding motifs.

Presentation

Anti-G3BP1 (mouse monoclonal),. One vial containing 50 µg of protein G purified mouse monoclonal IgG1ĸ in buffer containing 0.1 M Tris-glycine, 0.150 M NaCl, pH 7.4, and 0.05% sodium azide before the addition of 30% glycerol. Store at -20°C .
Normal Mouse IgG, . One vial containing 125 µg of mouse IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C.
RIP Primers, c-myc 3' UTR. One vial containing 75 μL of 5 μM of each primer specific for human c-myc 3' UTR. Store at -20°C.
FOR: CAC AAA CTT GAA CAG CTA CGG
REV: GGT GAT TGC TCA GGA CAT TTC 

Key Applications

• Western Blotting
• RNA Binding Protein Immunoprecipitation (RIP)

注意事项：

Application Notes

Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from 293 cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG, (Cat. #CS200621), or 5 µg of Anti-G3BP1 antibody (Cat. # CS204311). Ten percent of the precipitated proteins (lane 1: mouse IgG, lane 2: G3BP1) and 293 whole cell lysate (lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with Anti-G3BP1 antibody (Cat. # CS204311, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) 
Arrow indicates G3BP1.
Western Blot Analysis: 
Representative lot data.
K562 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-G3BP1 (0.1 ug/mL dilution). 
Proteins were visualized using a Goat Anti-Mouse antibody conjugated to HRP using a chemiluminescence detection system. 
Arrow indicates G3BP1 (~52 kDa).

Storage Conditions

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. 
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.